Universal rapid diagnostic test reader with trans-visual sensitivity

ABSTRACT

A universal and standalone rapid diagnostics test reader is disclosed herein that includes: a set of control electronics, an illumination component, an imaging component, a housing component, a wireless communication component, a rapid diagnostics test component, a universal rapid diagnostics test tray, wherein the tray can hold at least one rapid diagnostics test component having a shape and a size in a fixed position relative to the imaging component and the illumination component, and wherein the reader can accommodate more than one different rapid diagnostics test component, while using the same universal rapid diagnostics test tray. A universal rapid diagnostics test tray for a reader is also disclosed that includes: a rapid diagnostics test component; a tray component that is designed to operatively couple with the reader, wherein the reader can analyze more than one different rapid diagnostics test components, while using the same universal rapid diagnostics test tray; and a security component, wherein the security component operatively secures the rapid diagnostics test component in place on the tray component, wherein the tray component can hold the at least one rapid diagnostics test component having a shape and a size in a fixed position relative to the imaging component and the illumination component.

This United States Continuation in Part application claims priority toU.S. Provisional Application No. 62/362,538, entitled “Universal ELISAReader” and filed on Jul. 14, 2016; U.S. Utility application Ser. No.14/313,615, which claims priority to U.S. Provisional Application Ser.No. 61/889,821 entitled “Universal Rapid Test Reader for Lateral FlowImmunoassays with High Sensitivity”; U.S. Provisional Application Ser.No. 61/845,742 entitled “Flash-controlled, Wireless, Lensless, UniversalRapid Diagnostics Test (RDT) Reader”; U.S. Provisional Application Ser.No. 61/899,116 entitled “Non-uniform Flash Illumination Based focusingMethod for the Imaging of Targets that are uniformly illuminated”; U.S.Provisional Application Ser. No. 61/852,691 entitled “Flash illuminationbased Universal Rapid Diagnostics Test Reader running on a Cell-phone”;and U.S. Utility application Ser. No. 15/430,698 entitled “DiagnosticTesting Assays and Related Devices with Security and Methods of UseThereof” filed on Feb. 13, 2017, all of which are commonly-owned andincorporated herein by reference in their entirety.

FIELD OF THE SUBJECT MATTER

The current application is related to various embodiments of a universalrapid diagnostic test reader with trans-visual sensitivity.

BACKGROUND

Rapid diagnostic tests (RDTs) play an important and growing role in thecontinuum of care worldwide. Administered either at the point of care indoctors' offices, hospitals, urban and remote clinics, or by ambulatoryhealth workers and providing immediate results these tests contribute toimproved access, lower cost, and better-quality healthcare. Anincreasing number of RDTs are available for home use by patients and thegeneral public for testing of acute and chronic conditions. The dominanttechnology used for RDTs is Lateral Flow Immuno-Chromatographic assay(LFI) and with the worldwide annual value of LFI tests and services of$18B according to BCC Research. RDTs are also available in othervariations of immunoassays, such as fluorescent LFIs, flow-through, anddipstick tests. In fact, contemplated embodiments described here areapplicable to any RDT using a change of the optical properties as themechanism of action.

As valuable as RDTs are, they can be less reliable and accurate, becausethey are typically read visually, and therefore, are subject to humanerror [1-19]. These inherent errors can be substantially alleviatedthrough the use of electronic readers originally developed by ESE GmBHand today available from a number of sources [21, 22]. They aretypically desktop instruments for laboratory use, can be rather largeand heavy and can cost thousands of dollars. Recently, significantprogress in the state-of-the-art technology was achieved by ProfessorAydogan Ozcan and his research group at UCLA using a smartphone as thetechnology platform. In addition, they developed a reader [17, 20](hereafter Mudanyali reader) with the following advantages: a) small,handheld and light (˜2.3 oz), b) sensitive and accurate withtransmission or reflection readout mode, c) impervious to ambientlighting conditions, d) automated test readout with electronic datacapture and telemetry using smartphone communication capabilities, e)centralized data collection with geomapping capabilities and interfacesto health information systems, and f) low cost achieved by piggybackingon the enormous production volume of smartphones.

Despite the advantages, there are opportunities available for theseconventional readers to be improved. For instance, conventional systemsachieve low cost by using a smartphone which is inserted into a readerbody that provides RDT illumination, ambient isolation, and cassettehousing. However, different models of smartphones from a singlemanufacturer or even more from a variety of vendors all have differentmechanical dimensions, and they wouldn't fit into a body designed forone specific smartphone model. This precludes users from using their ownsmartphone for the reader: they have to buy another dedicated smartphonewhich is a significant cost increase, not to mention adding to thenumber of devices that need to be stored and utilized.

Readers require sources of illumination and associated controlelectronics and at least one battery housed outside the smartphone. InMudanyali's conventional reader, the control is provided by the softwareapplication in the smartphone via a physical cable, which plugs into thesmartphone micro USB power connector. External cabling adds to the costand reduces reliability; besides, many smartphones do not have thecapability for outbound control through their power connector. Also, thereference that discloses the Mudanyali reader describes a power sourcedisposed in the attachment, such that the self-powered reader can becontrolled via a physical button located on the attachment. Thisoperation fully depends on operator's ability to use the reader andincreases the complexity of operation. It would be ideal if contemplatedreaders and systems corrected many of the before mentioned deficienciesof the prior art, such as by simplifying communications to and from thereader, minimizing or removing additional hardware or systemrequirements, providing a universal test tray, test port, or testinsertion system, so that the reader and related system can be utilizedfor as many tests as possible without modification. It would also beideal if contemplated readers were simplified altogether by removing thenecessity for separate control by or use of a separate wireless device,such as a smartphone, tablet, or any other wirelessly-enabled device.

Moreover and more importantly with respect to contemplated embodimentsherein, Mudanyali's reader is capable of accommodating different teststypes using special customized-trays per cassette type, but thesecustomized trays must be changed out for each type of test cassetteused. Therefore, it doesn't provide a universal solution to image anytest without additional mechanical components. This requiredcustomization can increase the initial expense of ownership and use,along with leading to a system that contains more “moving pieces” orcomponents that need to be stored and utilized. A user must be able toknow which customized tray to use and must properly store it, so thatanother user can find the customized tray and know how it should be usedwith respect to the individual test. A universal reader should bereadily able to work with a significant number of different testcassettes without a need for any mechanical adaptation or additionalmechanical components, unlike the ones described above.

Recently another implementation of a smartphone-based reader has beendisclosed [23], which depends on the optimized Rayleigh/Mie scatterdetection by taking into consideration the optical nitrocellulosemembrane and gold nanoparticles on rapid tests. For each test type, thisapproach requires a complicated and precise calibration procedure todetermine the optimum angles of illumination that minimize the Miescattering from the membrane while maximizing the Rayleigh scatterdetection from the gold nanoparticles on and inside the membrane.Because of the significant variation between different RDT types andalso the variation within the samples of same RDT type in terms of useof components (e.g., membranes and nanoparticles) andposition/orientation of membrane and cassettes, successfulimplementation of this concept on a portable unit is quite challengingand not feasible. For instance, the coefficient of variation (CV)exceeds 50% in some of their measurements on quantitative tests [23].This reader variation is generally not acceptable even in qualitativemeasurements. This alignment-dependent approach may be useful forresearch purposes on the analysis of custom-made immunoassays usingadvanced optical imaging setups that includes a precise automatedscanning stage and other opto-mechanical components, but it is not theright approach for hand-held, real-time rapid diagnostic tests.

Note that although the work here was focused onsmartphone/wirelessly-enabled device-based andsmartphone/wirelessly-enabled device-activated/managed RDT readers asthe most advantageous architecture, many of the technologies describedherein apply equally well to any reader architecture based on digitalimaging.

SUMMARY OF THE SUBJECT MATTER

A universal and standalone rapid diagnostics test reader is disclosedherein that includes: a set of control electronics, an illuminationcomponent, an imaging component, a housing component, a wirelesscommunication component, a rapid diagnostics test component, a universalrapid diagnostics test tray, wherein the tray can hold at least onerapid diagnostics test component having a shape and a size in a fixedposition relative to the imaging component and the illuminationcomponent, and wherein the reader can accommodate more than onedifferent rapid diagnostics test component, while using the sameuniversal rapid diagnostics test tray. In some embodiments, the at leastone rapid diagnostics test component or test cassette comprises at leastone microplate, microtiter, ELISA plate, at least one micro-array assay,at least one biological and/or chemical test, at least one other assay,or a combination thereof.

A universal and standalone rapid diagnostics test reader is disclosedherein that includes: a set of control electronics, an illuminationcomponent, an imaging component, a housing component, a wirelesscommunication component, a rapid diagnostics test component, a displaycomponent, a universal rapid diagnostics test tray, wherein the tray canhold at least one rapid diagnostics test component having a shape and asize in a fixed position relative to the imaging component and theillumination component, and wherein the reader can accommodate more thanone different rapid diagnostics test component, while using the sameuniversal rapid diagnostics test tray. In these embodiments, the displaycomponent is physically integrated into the reader.

A universal rapid diagnostics test tray for a reader is also disclosedthat includes: a rapid diagnostics test component; a tray component thatis designed to operatively couple with the reader, wherein the readercan analyze more than one different rapid diagnostics test components,while using the same universal rapid diagnostics test tray; and asecurity component, wherein the security component operatively securesthe rapid diagnostics test component in place on the tray component,wherein the tray component can hold the at least one rapid diagnosticstest component having a shape and a size in a fixed position relative tothe imaging component and the illumination component.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1A shows Universal RDT reader attachment that can repeatedlyattached/detached to a cell-phone.

FIG. 1B shows a PCB is enclosed within the reader attachment.

FIGS. 2A and 2B show Different schematic views of proposed universal RDTreader prototype installed on an Android phone (Motorola Defy XT 535).This compact attachment can be repeatedly attached/detached to thesmartphone body without the need for fine alignment and anymodification.

FIG. 2C shows a Mechanical body of the reader attachment will alsoensure the isolation to the RDTs that are loaded to the smartphonereader attachment using a universal test tray. With its slip-free gripdesign, it allows the user to conveniently hold it even in the fieldsettings. It includes a single USB port with a photo-sensor tocommunicate with both smartphone and the PCB board. Having anunconventionally large field-of-view of ˜45 mm×85 mm, this attachmentwill also allow the user to acquire images of other objects of interest,such as user ID card (e.g., military) and RDT pouch with type/lotnumbers, while the universal RDT tray is retracted from the baseattachment assembly.

FIG. 2D shows a Reader attachment uses inexpensive optical components,i.e., a plano-convex lens (optional) and multiple diffused narrow-bandLEDs embedded on a single PCB that also carries circuitry for the dataand power communication with the smartphone and rechargeable flat or AAbatteries, respectively.

FIG. 2E shows a Schematic diagram of the proposed optical geometry isshown. (f) Flexible lens assembly is demonstrated. Utilizing an externaldiffuser and a narrow band-pass (<30 nm) filter, flash light of thesmartphone can be used to potentially support or mimic the LEDillumination.

Without any modification on the architecture of the main mechanical body(c), lens holder can be modified to hold the additional diffuser andband-pass filter as shown in FIG. 2F.

FIG. 3A shows a single universal RDT tray that will accommodate multiple(at least seven) RDT types.

FIG. 3B shows a contemplated single universal RDT tray that willaccommodate multiple (at least seven) RDT types.

FIG. 3C shows a contemplated single universal RDT tray that willaccommodate multiple (at least seven) RDT types.

FIG. 3D shows a contemplated single universal RDT tray that willaccommodate multiple (at least seven) RDT types.

FIG. 3E shows a contemplated single universal RDT tray that willaccommodate multiple (at least seven) RDT types.

FIG. 3F shows a contemplated single universal RDT tray that willaccommodate multiple (at least seven) RDT types.

FIG. 3G shows a contemplated single universal RDT tray that willaccommodate multiple (at least seven) RDT types.

FIG. 4 shows Rapid Test Reader prototypes fabricated using a 3-Dprinter. (a) A prototype with a slip-on universal RDT tray canaccommodate up to 7 different RDTs (b) A single-piece design that withrelatively smaller dimensions can accommodate and fully enclose up to 7different RDTs (c) A third single-piece design that can fully encloseand accommodate up to 5 different RDTs has significantly smallerdimensions and weight as well as smooth edges and corners. (d, e, f)Different types of RDTs, e.g., Uni-Gold™ HIV RDT (d), Cardiac Panel Test(e) and a custom hand held assay (f) can inserted to the prototype shownin (c). (g) and (h) Different views of the reader prototype shown in(c).

FIG. 5 shows views of the reader with its two-sided tray design for allfive different RDT types, including, (a) Afla-V aflatoxin RDT (70×27×8mm), (b) BioThreat Alert™ Anthrax RDT (62×30×6 mm), (c) Cardiac PanelRDT (80×32×5 mm), (d) a custom hand held assay (70×20×5 mm) and (e)Uni-Gold™ HIV RDT (Trinity Biotech) (39×18×6 mm). Accommodating threeout of five RDT types, the tray lid can be replaced with another one toaccommodate other RDT types without any modifications on the main bodyof the attachment.

FIG. 6 shows a view of the reader with a display of the main menu offunctionalities.

FIG. 7A shows one-design-fits-all universal cassette holder.

FIG. 7B shows universal Tray with a spring to accommodate any test thathas dimensions of 85 mm by 35 mm or smaller.

FIG. 7C shows top-view schematics of the tray (left) and side-viewschematics of the tray (right).

FIG. 8 shows reflection and transmission readout modes.

FIG. 9 describes that we have conducted initial measurements to test theperformance of the proposed optical imaging scheme and digital imageprocessing algorithm by analyzing a Bio Threat Alert™ RDT (TetracoreInc) activated using highly diluted positive control sample. In order tosimulate the proposed reader platform, we conducted this measurementwith a Samsung Galaxy smartphone mounted on an optical table withadjustable optical and mounting components. For the illumination of RDTunder test, two narrow-band LED arrays (wavelength=565 nm, bandwidth=˜30nm) were used. (a) Prior to acquire RDT image using the proposed opticalscheme, under ambient light, we have recorded a basic smartphone cameraimage in the room conditions, providing no distinct color intensity (onthe test line) that can be identified by human eye. However, in thedigitally processed and enhanced image of the same RDT acquired usingour optical platform (b), digital contrast level has been significantlyimproved between the test line and the background. (c) Although therehas been an analyte-generated background in this RDT image, ourderivative-based filter has corrected this issue, enabling the recoveryof test as well as control line free from the chemical and opticalbackground noise.

FIG. 10 shows exemplary (recorded and automatically processed) images ofnegative (bottom row) and positive (top row) tests for (a) Afla-Vaflatoxin RDT, (b) Cardiac Panel RDT, (c) a custom hand-held assaydeveloped by ECBC (Edgewood Chemical Biological Center), (d) BioThreatAlert™ Anthrax RDT, and (e) Uni-Gold™ HIV RDT (Trinity Biotech).

FIG. 11 shows the quantitative calibration curve for the Aflatoxin testthat was generated using 160 different measurements is shown. For eachconcentration level, 2 different RDT were imaged 5 times.

FIG. 12 shows the illumination pattern in the transmission mode

FIG. 13 shows the summary of our measurements on the dry standardOrasure RDT kits which represents various concentration levels rangingfrom blank (0) and negative (N) to high density levels (>5+). In thetable below, the measurement intensity mean, standard deviation and CVvalues are shown together with the corresponding optical density levelsprovided by Orasure.

FIG. 14 shows the schematics for the transmission imaging/readoutmodality using the cell-phone flash and a mirror.

FIG. 15 shows universal reader design that can work with any smartphoneor tablet, utilizing smartphone cases customized for each phone ortablet type.

FIG. 16 shows that, by the optimized selection of the common location offlash/camera for various phones and tablets, the universal readerattachment can work with various smartphones and tablets.

FIG. 17 describes the communication between the smartphone and tablet(application) and the embedded PCB. The reader application running onthe smartphone or tablet can control the flash pulses generated by thecamera and control the electronics on the PCB.

FIG. 18 shows various views of a wireless and standalone universal RDTreader that is enclosing all opto-electro-mechanical components and iscontrolled by at least one mobile device (smartphone, tablet, PC) viaBluetooth or similar connections as shown in (b)

FIG. 19 shows various views of a wireless and standalone universal RDTreader that is enclosing all opto-electro-mechanical components and iscontrolled by an embedded display/user interface. The same device can bealso controlled by at least one mobile device via Bluetooth or similarcomponents as well as shown in FIG. 18 (b).

FIG. 20 shows the internal components of a wireless and standaloneuniversal RDT reader, such as the one from FIGS. 18(a) and 19.

FIG. 21 shows a contemplated spring-loaded universal tray component thatcan be utilized in a contemplated universal RDT reader.

FIG. 22 shows a contemplated universal tray component comprising a “rackand pinion” design for use in or with a contemplated universal RDTreader.

DETAILED DESCRIPTION

A universal rapid diagnostics test reader is disclosed and describedherein that addresses many of the shortcomings of conventional readersand technology. Specifically, contemplated universal rapid diagnosticstest readers accomplish the following: a) compatible with smartphones,tablets, laptops, computers, and any other wirelesscommunication-enabled or included device, b) compatible with Android andIOS platforms, c) works well with any assay or test, because of auniversal tray design and modular software package, d) trans-visuallimit of detection (OD levels down to 0.2-0.3%), e) low measurementcoefficient of variation (down to 0.1%), f) battery powered andrechargeable via USB or another suitable power supply, g) operationbattery life up to 12 hours with a standby life of up to 24 hours (atthis time), h) wireless range for operation up to 100 meters, i)packaging is smaller than 175 mm×72 mm×72 mm, and j) weighs less than 2pounds.

Specifically, a contemplated universal and standalone rapid diagnosticstest reader is disclosed herein that includes: a set of controlelectronics, an illumination component, an imaging component, a housingcomponent, a wireless communication component, a rapid diagnostics testcomponent, a universal rapid diagnostics test tray, wherein the tray canhold at least one rapid diagnostics test component having a shape and asize in a fixed position relative to the imaging component and theillumination component, and wherein the reader can accommodate more thanone different rapid diagnostics test component, while using the sameuniversal rapid diagnostics test tray. In some embodiments, the at leastone rapid diagnostics test component or test cassette comprises at leastone microplate, at least one microtiter plate, at least one ELISA plate,at least one micro-array assay, at least one biological and/or chemicaltest, at least one other assay, or a combination thereof.

Another contemplated universal and standalone rapid diagnostics testreader is disclosed herein that includes: a set of control electronics,an illumination component, an imaging component, a housing component, awireless communication component, a rapid diagnostics test component, adisplay component, a universal rapid diagnostics test tray, wherein thetray can hold at least one rapid diagnostics test component having ashape and a size in a fixed position relative to the imaging componentand the illumination component, and wherein the reader can accommodatemore than one different rapid diagnostics test component, while usingthe same universal rapid diagnostics test tray. In these embodiments,the display component is physically integrated into the reader.

A contemplated housing component is designed to enclose all componentsof a contemplated reader into a light tight enclosure and wherein therapid diagnostic test component is illuminated only by the illuminationfrom the reader and not illuminated by ambient light. This concept willbe additionally discussed herein.

As used herein, “rapid diagnostics test component” or “test cassette”can be used interchangeably and can include at least one microtiterplate, at least one microplate, at least one ELISA plate, at least onemicro-array assay, at least one biological-chemical test, or at leastone other assay (immuno-assays or others with spatial features toindicate certain conditions). Other suitable rapid diagnostic tests canalso be used, as long as they are designed to fit into a tray componentarchitecture and be analyzed by a reader-type device, such as thosecontemplated herein.

Many of the deficiencies as outlined earlier are corrected by thecontemplated embodiments disclosed herein. Specifically, contemplatedembodiments overcome the following limitations:

-   -   Different cassette and test component designs and mechanical        dimensions make it challenging to interface with the imaging        system of the reader. Contemplated embodiments work with a wide        range of cassette or assay sizes and types, and they avoid the        pitfalls of mechanical adapters for each test type.    -   Conventional readers require sources of illumination and        associated control electronics and battery housed outside the        smartphone. In Mudanyali's conventional reader, the control is        provided by the software application in the smartphone via a        cable which plugs into the smartphone micro USB power connector        or a physical switch that is outside the attachment. External        cabling and physical switches add to the cost and reduce        reliability; besides, many smartphones do not have the        capability for outbound control through their power connector.        Current contemplated embodiments solve this problem with a        wireless control (no wires, cables or physical switches needed).    -   Conventional systems achieve low cost by using a smartphone        which is inserted into a reader body that provides RDT        illumination, ambient isolation, and cassette housing. However,        different models of smartphones from a single manufacturer or        even more from a variety of vendors all have different        mechanical dimensions, and they wouldn't fit into a body        designed for one specific smartphone model, which precludes        users from using their own smartphone for the reader: they have        to buy another dedicated smartphone which is a significant cost        increase. Current embodiments provide a low cost way to        eliminate this problem and enable the use of a wide variety of        mobile devices including smartphones and tablet PCs.

Architecture

A Universal Rapid Diagnostics Test (RDT) Reader has been developed, isdisclosed herein and a contemplated embodiment is shown in FIG. 1A, thatcan accommodate multiple (at least 5 different test types withdimensions up to 35 mm×85 mm) RDTs without any need for mechanicalmodification or external RDT trays. FIG. 1A shows a contemplateduniversal RDT reader attachment 110 in an open 115 and closed 120position. A contemplated communications device, such as a cell phone, isnot shown in this Figure. In this single-piece adapter design, a printedcircuit board (PCB) 150, as shown in FIG. 1B is utilized that includesmultiple illumination light emitting diodes (LEDs), a replaceable andrechargeable battery, a recharging circuit with its USB port, and aphoto-sensor, which is used to wirelessly trigger/control theillumination LEDs via the cell-phone application. The PCB with thebattery is coupled with or affixed into the adapter. The complete readerattachment assembly is wirelessly press-fit onto a smartphone for easymounting and demounting.

One objective of the development of some of the contemplated embodimentsdisclosed herein was to introduce a rugged, wireless (controlled, insome embodiments, using cellphone flash via the photo-sensor), lensless(no external lens needed), smartphone based universal RDT reader thatcan continuously operate over extended hours even in field settings. Itshould be understood that the platform is cell-phone or wireless-deviceindependent, such that it can be adapted to any cell-phone device orwireless device with minor or no mechanical modifications. In thisembodiment, we used an inexpensive and rugged Motorola Defy XT 535smartphone. It includes optical and electrical components embedded on aprinted circuit board (PCB) that is powered by a rechargeable battery,which can operate over 12 hours without any need for external power.This contemplated universal reader, without any modification on itsmechanical architecture, accommodates and digitally interprets a broadrange of RDTs to diagnose chemical and biological threats and otherdiseases.

In some embodiments, the universal reader may not need to have theillumination component, the imaging component, or any other componentcontrolled by a separate wirelessly-enabled device. The ability tooperate the control electronics component or any other component of theuniversal reader may be physically incorporated into the reader. Inthese instances, the universal reader still contains a wirelesscommunication component, or possibly a port for a wired connection, thatallows the data from the universal reader to be transferred securely toanother device, such as a smartphone, a tablet, a laptop, a computer, oranother suitable device. It should be understood, however, that theuniversal reader has a certain amount of memory and storage available,so that data does not need to be immediately transferred, but can bestored securely until the user is in a position to transfer the data foruse. This embodiment is shown in FIG. 19, which is described in detailherein. In some embodiments, a set of control electronics compriseprocessor electronics that control the illumination component, theimaging component, the wireless communication component, or acombination thereof.

In some embodiments, a contemplated illumination component comprises atleast one light emitting diode. A contemplated illumination componentincludes illumination of the rapid diagnostics test component by the atleast one light emitting diode at the wavelength of imaging forchromatographic and colorimetric rapid diagnostic test components or atthe excitation wavelength for the fluorescent rapid diagnostic testcomponents. In some embodiments, a contemplated illumination componentcomprises a reflection mode of operation, wherein at least onelight-emitting diode and the imaging component are on a front side ofthe rapid diagnostic test, and wherein the light-emitting diode axis isroughly perpendicular to the rapid diagnostic test component plane. Inother embodiments, a contemplated illumination component comprises atransmission mode of operation, and wherein the rapid diagnostic testcomponent is between the at least one light-emitting diode and theimaging component.

The sensitivity and accuracy of the platform was demonstrated byconducting repeated measurements on positive (including the onesactivated by highly diluted positive control samples) and negativetests. Through a custom-developed smartphone application, a contemplatedintegrated smartphone-based reader labels digitally processed testresults with spatiotemporal information and transfers them to centraldata collection points (servers) that can be accessed locally andglobally. In contemplated embodiments wherein the reader wirelesslycommunicates with the wirelessly-enabled device, such as a smartphone,tablet, or other device, data and test results are transferred to thoseplatforms, which may hold or control the central data collection point(locally or on the Cloud) or may transfer that data to another centraldata collection point.

In this particular embodiment, the smartphone-based RDT reader andspatiotemporal threat/disease monitoring platform utilizes a compactsnap-on smartphone attachment 210 that can be repeatedlyattached/detached at the back 295 of the smartphone devices 290 (seeFIGS. 2A and 2B) to acquire digital images of RDTs. This scalableattachment is designed to fit different smartphone devices (e.g., iPhoneor Android phones and tablets) and can be simply adapted with minimal orno engineering.

The mechanical body of this independent reader attachment is designed tobe robust and easy-to-handle by the user and initially prototyped usinga 3-D printer, which uses ABSplus™ modeling material, a recyclable andeco-friendly thermoplastic. For volume manufacturing using differenttechniques (e.g., injection molding or casting), other material typeswith different material properties can be used. This snap-on readerattachment utilizes inexpensive optical components and printed circuitboards with various electrical components i.e., multiple LEDs (lightemitting diodes) and/or LED arrays, a photo-sensor interface towirelessly control and trigger the illumination LEDs (via the cell-phoneapplication), rechargeable battery as well as a recharging circuitry andits USB port embedded on the same. (see FIGS. 2C and 2D). In FIG. 2C, acontemplated device 210 is shown with a USB connector 220, a “slip-free”grip design 230, a lens assembly 240, protection glass 250 and an “errorproof” multi-track system 260. In FIG. 2D, the back of a contemplateddevice 210 is shown with the USB port or connector 220, a rechargingcircuit 221, a battery holder 222, a LED driver circuit 223, an audioport 224 and at least one LED bar 225. Please note that this readerattachment does not require the use of any external lenses for themagnification/demagnification of the RDT image, however it has anoptional tray that allows the user to utilize an additional imaging lens(optional) to meet to requirements of different imaging conditions.

Another important design consideration is the choice of RDT illuminationscheme that has a significant effect on the sensitivity and accuracy ofthe test interpretation. Embedded on a single PC board, multiplediffused LEDs can be used to illuminate (with an illumination angleclose to normal incidence) the RDT under test (see FIG. 2D) in bothreflection and transmission modes (butt-coupled to the RDT under test).The PC board can also a high-power UV led for the digital evaluation offluorescent tests. Unlike reflection and transmission mode LEDs, UV ledis positioned with angle of ˜30-50 degrees to excite the tests ofinterest. Powered by an embedded rechargeable battery, LEDs arewirelessly controlled by a photo-sensor mounted on the PCB. Thesmartphone application, based on the user's selection, digitallyconfigures and utilizes the flash of the cell-phone to trigger thephoto-sensor and control the illumination LEDs automatically duringtesting. This wireless and digital communication with the illuminationLEDs allows the software application to turn on the LED arrays onlyduring image acquisition and observation, decreasing the powerconsumption for extended battery life.

In contemplated embodiments where the universal reader utilizes awirelessly-enabled device that is not physically connected or coupledwith the reader, there is an illumination component and imagingcomponent that are built into the reader. In these instances, theillumination component, the imaging component, or a combination thereofmay be controlled by the control electronics, the wirelessly-enableddevice, or a combination thereof.

Furthermore, as shown in FIG. 2D, the PCB board also has acustom-designed circuitry with an integrated USB port or connector torecharge the battery of the reader attachment without any need foradditional equipment, simply by sharing the smartphone USB cable withinthe base mechanical body. Moreover, by using this external battery, acontemplated RDT reader is able to evaluate >1000 RDTs and operatelonger than 12 hours without any need for external battery andrecharging. Applying<10 second exposure for image acquisition per test,this enables rapid readout of stacked RDTs without an interruption.

Rather than using broad-band light sources (e.g. ambient light) withvarying intensity profiles and optical spectrum, the use of special LEDillumination can significantly increase the contrast between thecontrol/test lines and the background on RDT images, provided that thewavelength of the illumination is optimized based on spectralmeasurements. For instance, test pads (e.g., nitrocellulose membrane) ofmost commercially-available RDTs produce distinct color signal by theimmobilization of colloidal gold-labeled antigen (e.g., analyte) andantibody (e.g., binding proteins) complexes, exhibiting similar spectralproperties. Based on our initial tests on various RDTs in the market,the use of LEDs with a peak wavelength of between 520 and 590 nmprovides the highest contrast if colloidal gold nano-particles are usedto label target antigen or antibody (in both reflection and transmissionmodes). It should be noted that this optimum center wavelength wasdetermined based on spectral measurements on colloidal gold-based RDTsthat are widely commercially available. Together with thecustom-developed image processing algorithm that is discussed later,optimization of the illumination wavelength has primary importance onthe trans-visual sensitivity of this smartphone based reader platform.

In parallel to the proposed LED illumination scheme, the use of theflash light was examined as an additional illumination source that isalready available in the camera smartphone devices (see FIG. 2F). FIG.2F shows a lens assembly 240 wherein two different systems aredisclosed: a flash system 242 having a diffuser 243 and a lens 244 or aLED system 246 having a lens 244, whereby one of these systems isinstalled in the lens assembly 240. Locating a band-pass filter(optional) that is butt-coupled to the flash light, flash can bealternative to the LED illumination that has been described above (inboth reflection and transmission modes). Although the use of externalLEDs has significant advantages, such as high-power output, opticaldesign flexibility and illumination uniformity, implementing the cameraflash light on the RDT reader, as an option for less demandingapplications, can eliminate the need for using LEDs, electricalcomponents, and external battery. In reflection mode, by simply locatinga band-pass filter (e.g., with 520-590 nm pass-band), the flash lightcan replace the reflection mode LEDs. On the other hand, flash light canbe also used for the transmission mode imaging. A reflection elementsuch as a mirror can be placed at the top wall of the attachment toreflect the image of the RDT that is positioned in front of the flashfor transmission mode imaging. Reflected image of the RDT can be thenimaged by the cell phone camera (see FIG. 14).

In another contemplated embodiment that is shown in FIG. 18 and FIG. 19,a wireless or USB based reader platform (1801 and 1901) to digitallyimage and analyze microtiter plates, microplates, ELISA plates,micro-array assays, biological-chemical tests, and other assays(immuno-assays or others with spatial features to indicate certainconditions). The rapid diagnostic test components are not shown in theseFigures, but instead is shown in other Figures, such as FIG. 21(reference number 2110).

FIG. 19 shows a contemplated reader that is controlled using an embeddeduser display 1902 or by a wirelessly-enabled mobile device 1805 (shownin FIG. 18). These contemplated embodiments shown in FIGS. 18 and 19utilize an image analysis software/application and an opto-electronicimaging box to digitally record images of assays and transmit the imagesto a remote or local device that is wirelessly-enabled (phone, tablet,computer, server) 1805 using its already integrated wirelesscommunication modules (WIFI, Bluetooth, WIFI direct, infrared, etc.)and/or USB port via USB cable. In addition to the remote or localdevices, it can be also controlled and operated via an embedded displaywhich serves as a user interface 1902. The image of the assay (digitallytransferred from imaging box to remote/local device) is processed byimage analysis software/application for the digital, accurate andquantitative (colorimetric, luminescent, fluorescent andchromatographic) assay interpretation.

As shown in FIG. 20, a contemplated imaging box can utilize single ormultiple digital sensors (e.g., CMOS or CCD) 2002 to digitally recordthe single or multiple images of the assay under test. With thiscontemplated imaging module or component 2002, one or more digitalsensors and one or more illumination LEDs that include reflection andmultiple fluorescent modes, along with other optical components. In caseof the use of multiple cameras, the reader software can register theimages to create a single image. It can also use single or multipleinterchangeable LED arrays for illumination in transmission 2004 orreflection or fluorescent modes 2002.

FIG. 19 shows a side and internal view of a wireless and standaloneuniversal RDT reader with an embedded display/user interface. Thisreader can be also operated by a mobile device wirelessly. FIG. 20 showsthe internal components of a wireless and standalone universal RDTreader, such as the one from FIGS. 18 and 19.

The opto-electronic imaging box (FIGS. 18 and 19) includes at least oneof the following components; GPU, CPU, Flash Memory, RAM, USB andEthernet ports, GPIO pins for a data acquisition and processing 2001with a camera/imaging device 2002, Wireless (WIFI, Bluetooth, WIFIdirect, infrared, etc) communication module(s) and USB port for datacommunication, various sensors (e.g., motion sensor and light sensor),circuit board carrying all optical illumination sources, opticalcomponents (lenses and filters), and an imaging chamber as well as anoptional display as a user interface. The imaging box or universalreader is powered by an integrated battery 2003, which can be recharged.The Application Specific Integrated Board (ASIB) 2001 carries orincludes one or more of the following components; CPU, GPU, FlashMemory, SD Card slot, USB ports, Ethernet ports, GPIO pins, WiFi andBluetooth modules.

The tests are inserted to the reader/imaging box from the side using thetray 1804. The imaging box can be controlled by the embedded display1902 or mobile device (wirelessly or using a USB cable) 1805. After thetarget (test) is inserted, the reader box configures the opto-electroniccomponents and turns on the optical illumination sources (LEDs, lasers,etc.), acquires the image or video of the target, and then transmit thisdata to the reader software/application running on a phone, tablet, pc,laptop, or server (for remote computation and analysis). The data can bealso presented on the embedded display (optional) 1902. The readersoftware can be programmed and configured to analyze the images of anyassay (immuno-assays, lateral flow tests, flow through tests, etc.). Itpresents the images (acquired by the reader box) and finalinterpretation as well as test results to the user based on initialcalibration.

Users can install the reader application and use their own mobile device1805 to connect and control the instrument wirelessly, or can use a USBcable attached to the system to operate various aspects of acontemplated universal reader, however, it should be understood that oneof the advantages of contemplated readers and systems are that the useris unencumbered by cords, wires, or multiple trays. A contemplatedreader and/or system can work as a part of cloud or provide astand-alone reader solution. As an alternative, a contemplated systemcan be operated and controlled using the optional embedded display. Inthis case, wireless and remote devices can be still used.

Cassette Tray or Tray Component

Enclosing the optical imaging interface, the mechanical body of thereader attachment will also ensure the isolation to the RDTs that areloaded to the smartphone reader attachment as shown in FIGS. 1 and 3.Prior to digital evaluation, an RDT is press-fit into a unique cradle onthe tray in an easy, reliable, and error-proof operation. On theinterior side of the universal tray, a series of cradles are formed byridges at different heights, orientations and lateral extensions, asshown in detail in FIG. 3, for 7 different RDTs. A computer-aidedoptimization and design methodology used to design this universal trayis akin to solving a three-dimensional puzzle and can be readily appliedto a different set of RDT shapes and sizes.

Without any modification on the base reader attachment shown in FIG. 2E,multiple trays can be used that are capable of holding at least 5 kindsof RDTs each. FIG. 2E shows a contemplated device 210 where a smartphone290 is connected having a CMOS or complementary metal-oxidesemiconductor 292 and a lens 294. An additional lens 245 is shown, withthe protective glass 250 and the smart track or multi-track system 260.The PCB 275 is also shown. A RDT tray 280 and the tray lid 285 is alsoshown. As a matter of fact, this innovative design eliminates the needof using an individual customized tray for each RDT type andsignificantly decreases the material cost and logistical problems,enabling ease-of-use even in the field settings. By the optimization ofthis innovative approach, the mechanical interface (tray) can bedesigned to accommodate different and larger groups of RDTs. It is alsoimportant to underline that this universal tray fully encloses the RDTsof interest (see FIG. 2A-F) to tackle potential ambient light leakageinto the optical attachment. Since RDT material/packaging may behave asa waveguide that couples the ambient light to the optical imaginginterface, this universal tray design is a vital design feature toensure the repeatability of the measurements.

Moreover, the proposed RDT reader attachment will have a physicalopening (i.e., field-of-view) of ˜45 mm×85 mm to accommodate thisuniversal tray carrying a wide range of RDTs (see FIG. 3A-G). FIGS. 3Athrough 3G show various RDTs 330 that are coupled with the RDT readerattachment 310, wherein the RDT reader attachment is shown in a sideview (upper view) and an above perspective (lower view) in each Figure.Users can rapidly replace an already evaluated RDT with a new one to betested without having any mechanical difficulty, thus enabling testingof large number of stacked RDTs in a short time.

Also, it also allows the user to acquire images of other objects ofinterest, such as user ID card and RDT pouch with type/lot numbers,while the universal RDT tray is retracted from the base attachmentassembly. Digitally linked to the test results, these additional imagescan be processed to extract the relevant identification and securityinformation. Note that the unconventionally wide field-of-viewintroduced here to accommodate a broad range of RDTs provides anopportunity to acquire images of even larger RDTs with larger dimensionsor non-planar packaging (i.e. urine cup) by partially sacrificing thecompactness of the reader attachment. An embodiment of these designprinciples is shown in FIGS. 4-6.

FIG. 4 shows (a, b, c) Photo of the Rapid Test Reader prototypes 410fabricated using a 3-D printer. (a) A prototype with a slip-on universalRDT tray can accommodate up to 7 different RDTs (b) A single-piecedesign that with relatively smaller dimensions can accommodate and fullyenclose up to 7 different RDTs (c) A third single-piece design that canfully enclose and accommodate up to 5 different RDTs has significantlysmaller dimensions and weight as well as smooth edges and corners. (d,e, f) Different types of RDTs, e.g., Uni-Gold™ HIV RDT (d), CardiacPanel Test (e) and a custom hand-held assay (f) can inserted to theprototype shown in (c). (g) and (h) Different views of the readerprototype shown in (c).

FIG. 5 shows views of the reader 510 in an open embodiment showing itstwo-sided tray design for all five different RDT types 530, including,(a) Afla-V aflatoxin RDT (70×27×8 mm), (b) BioThreat Alert™ Anthrax RDT(62×30×6 mm), (c) Cardiac Panel RDT (80×32×5 mm), (d) a custom hand-heldassay (70×20×5 mm) and (e) Uni-Gold™ HIV RDT (Trinity Biotech) (39×18×6mm). Accommodating three out of five RDT types, the tray lid can bereplaced with another one to accommodate other RDT types without anymodifications on the main body of the attachment.

FIG. 6 shows a view of the reader 610 with a display of the main menu offunctionalities as shown on the smartphone 690 attached.

An alternative contemplated design for a universal cassette holder 710is shown in FIG. 7A. In this contemplated embodiment, the position ofthe cassette 730 is fixed by pushing it against an L-shaped ridge 720;the cassette is kept in place by one or more leaf springs 725. Byleaving the two sides of the cassette unconstrained, this design canaccommodate a wide variety of shapes and sizes with a single design.This kind of flexibility is a huge advantage for the manufacturer andthe users. Rather than just pushing the cassette into the only cradle itwould fit, now the operator must make sure that the cassette fits snuglyin the corner of the L. Also, for a triangular or other non-rectangularshapes, the test strip is at an angle but this is easily handled by thesoftware (application running on the cell-phone). Moreover, thisflexible approach, without any hardware and software modifications, mayallow the user to work with emerging next-generation technologies (flowthrough tests) that will be available at the markets in the near future.

In addition to the designs shown in FIG. 1A and FIG. 7A, a completelyuniversal and user-friendly tray design 710 has been developed forcontemplated readers (FIG. 7B). The designs are based on the conceptsshown in FIG. 7C: by constraining the position of the RDT 730 in oneplane only by an L-corner 720 and in the perpendicular direction by aslanted or curved side (727 in FIG. 7C) it is possible to accommodate awide range of RDT shapes and sizes in the same tray. This designutilizes another contemplated flat tray with a spring 725 placed on theside wall to keep the cassettes, which can simply slide into the tray,in place. In this design, the tray has only three walls with an openingat the top for the user to slide in the test cassettes of interest. Theposition of the cassette is fixed by pushing it against to the side ofthe tray; the cassette is kept in place by one or more leaf springs.This design allows the user to work with any cassette type that issmaller than 85 mm by 35 mm. By simply modifying the tray (testinterface, not the main body of the attachment), these dimensions can befurther increased.

On the other hand, operator inserts the cassettes into this mechanicalinterface by sliding them in all the way into the tray such that ideallythe cassette should fit in the L-corner. However, unlike the drawback ofthe design in FIG. 7A, the potential shift due to the failure of theoperator will be only in longitudinal direction—not in other directions.Our smart cell-phone application will recognize such vertical shifts onthe position of test casettes and either digitally compensate for it andevaluate the test or warn the operator to correct the position of thetest cassette.

Alternatively, a second spring could be implemented on the fourth sideof the tray to ensure positive contact with the L-corner. To keep thecassette from falling out of its position, a third spring or springscould be mounted on the top of RDT. Alternatively, at least one of thetray sidewalls could be slanted 727 or curved as shown in FIG. 7C; notethat different cassette thicknesses 730 all fit in with only a slightlateral displacement. In FIG. 7C, a top-view 702 and a correspondingside-view 704 is shown.

Methods of utilizing the readers and the tray components disclosedherein include: providing at least one first rapid diagnostics testcomponent having a first physical size, first feature and first format;providing at least one second rapid diagnostics test component having asecond physical size, second feature and second format; inserting thefirst rapid diagnostics test component in a contemplated universal rapiddiagnostics test reader; analyzing the first rapid diagnostics testcomponent using the universal rapid diagnostics test reader; removingthe first rapid diagnostics test component from the reader; insertingthe second rapid diagnostics test component in a universal rapiddiagnostics test reader without any mechanical adjustments of the readeror without the use of any additional parts or additional inserts; andanalyzing the second rapid diagnostics test component using theuniversal rapid diagnostics test reader.

A universal rapid diagnostics test tray for a reader is contemplatedthat includes: a rapid diagnostics test component; a tray component thatis designed to operatively couple with the reader, wherein the readercan analyze more than one different rapid diagnostics test components,while using the same universal rapid diagnostics test tray; and asecurity component, wherein the security component operatively securesthe rapid diagnostics test component in place on the tray component,wherein the tray component can hold the at least one rapid diagnosticstest component having a shape and a size in a fixed position relative tothe imaging component and the illumination component.

FIGS. 21 and 22 show additional contemplated tray designs that can beutilized with readers and imaging systems disclosed herein. FIG. 21shows several embodiments of a spring-loaded tray design, as alreadydisclosed herein. In this figure, a rapid diagnostics test tray 2101,comprises a spring 2120 and a rapid diagnostics test component 2110. Inthis Figure, it is clear that the spring-loaded design works well toprovide a secure holder to a number of different types of tests, testcomponents, and test designs.

FIG. 22 shows another embodiment of the universal tray design concept2201—this time using a rack and pinion design to hold various test andassay types in place securely in one universal tray. A contemplated testtray is shown from the top side or top view 2210 and the bottom side orbottom view 2220. A portion of the top side 2210 of a contemplated testtray 2205 is shown blown up to provide additional detail of the rack andpinion design. In this embodiment, rack 1 2230 and rack 2 2235 are usedto hold a rapid diagnostics test component (shown in the top side 2210embodiment) and those racks are adjusted through the use of a rotatablepinion 2240. The pinion 2240 engages with a slider 2260 and springs 2270to help to secure the test component. The bottom springs 2275 are shownin the bottom side view 2220. This embodiment also clearly shows anopening 2250 for transmission illumination, which is disclosed in detailherein.

Readout

A contemplated reader 810, like the conventional readers disclosedearlier, has three readout modes: fluorescent (not shown), reflectionmode 820 and transmission mode 830, as shown in FIG. 8. The reflectionmode 820 is used in all available readers as it obviously parallelsvisual readout, it is easy to implement, and provides comparableresults. FIG. 8 shows the cell phone camera 891, the cell phone cameralens placement 892, an external imaging lens 893, an HIV test membrane894 in a semi-transparent HIV cassette 896, as they are placed or asthey are relative to the reader 810. Conventional readers are used moreto avoid operator errors and obtain data in the electronic format ratherthan to improve the sensitivity. In the implementation, howevertrans-visual (better than visual) sensitivity was obtained even in thereflection mode by virtue of ambient isolation and optimization ofillumination, as described previously. A qualitative example 905 oftrans-visual performance is shown in FIG. 9 where the reader is clearlyable to detect a weak line invisible to the naked eye by using an imageprocessing algorithm similar to conventional readers. An optimized imagefor all five RDTs used in this work is shown in FIG. 10. FIG. 10 showsexemplary (recorded and automatically processed) images of negative(bottom row 1015) and positive (top row 1025) tests for (a) Afla-Vaflatoxin RDT, (b) Cardiac Panel RDT, (c) a custom hand held assaydeveloped by a third party, (d) BioThreat Alert™ Anthrax RDT, and (e)Uni-Gold™ HIV RDT (Trinity Biotech).

To demonstrate the accuracy of this reader in the reflection mode, astatistically significant number of tests were performed on one highquality RDT cassette capable of quantitative performance, Afla-Vaflatoxin RDT. The result is shown in FIG. 11. In reflection mode, inaddition to the at least one LED(s) located on the PCB, cellphone flashcan be used to illuminate the RDT under test.

Transmission mode readout was first proposed by Mudanyali et al [18] andqualitatively demonstrated to be an alternative to the reflection mode.However, their transmission mode required that both sides of the LFIstrip be open and accessible to light-one side toward the illuminatingsource and the other side toward the camera- and this turned out to be amajor impediment for practical use. The fact is that an overwhelmingmajority of RDT cassettes today on the market have only one open windowfor the strip with the other side being covered by a plastic back (forexamples see FIGS. 4 and 5), thus precluding the use of the transmissionmode except for a small number of specialized cassettes or cassettescustom designed for this mode.

The key insight that led to some of the contemplated embodimentsdisclosed herein is that nearly all RDT cassettes on the market are madeof white or lightly colored plastic that is sufficiently translucent toallow sufficient light transmission through the cassette wall to provideadequate illumination of the LFI strip and ultimately detection by thecamera. In addition, the translucent plastic acts as a diffusersubstantially improving the uniformity of the strip illumination. Acontemplated rapid diagnostic test component comprises a bare strip, astrip on a translucent plastic backing, a conventional strip packaged ina translucent plastic cassette, or a combination thereof.

The advantages of these contemplated embodiments are summarized asfollows:

-   -   The signal captured by the camera contains information about the        density of gold particles throughout the thickness        (3-dimensional morphology) of the paper strip rather than just        on the surface or close to the surface, which will substantially        increase the lower detectable limit compared with the reflection        mode measurement.    -   Translucent plastic in the cassette wall in the pathway of light        is generally strongly scattering and diffuses light which        contributes to the uniformity of the illumination of the strip        and this minimizes measurement errors.    -   The reflection mode illumination may cause multiple reflections        from the RDT cassette, causing trouble in digital processing        steps. This is particularly the case when the cassette window is        covered with plastic, which is the prevailing situation with        rapid tests used with saliva and which often also have weaker        signal. The transmission mode avoids these problems.    -   Because of the above all light reaching the camera is diffused        either through two layers of plastic with and without the strip        or one layer plus the strip. Consequently, the contrast over the        camera field of view is considerably more uniform than for the        reflection case as shown below, which reduces problems with        camera saturation and white balance variation.    -   For the same reason as above, there are no shadows around the        walls of the window (due to the oblique illumination angle of        the light sources in reflection mode) which are also detrimental        for the measurement. In fact, it was observed that the backside        illumination creates a sharp edge 1245 around the window which        may be beneficial in determination of the window position and        line edges (FIG. 12)    -   Blank (not-activated) tests (RDTs or LFIs) have test and control        bands. Antibodies or other chemicals that are necessary for the        color changes are dispensed (e.g., injected or coated) to these        bands and required for the successful operation. In transmission        mode, these bands can be screened before the use as a quality        control mechanism at the manufacturing side.

The performance of this transmission mode was confirmed using a set ofcalibration HIV RDTs. The results are shown in FIG. 13. They confirmtrans-visual sensitivity of <0.5% Optical Density and excellent CV of˜1%.

The transmission mode LEDs are located at the tray (behind the RDT undertesting), and coupled to the back of the plastic cassette. Moreover,instead of external LEDs, cell-phone flash can be used provided that amirror is located at the tray. In this flash-transmission geometry, theRDT is located on the top of the flash and parallel to the cell-phonecamera. The flash is controlled by the cell-phone application andilluminates the back of the cassette. The mirror located at the top ofthe tray reflects the image which is recorded via the cell-phone camera.See FIG. 14 for the transmission mode readout geometry using flash. InFIG. 14, the top view 1407 of a contemplated reader 1410 is shown intransmission readout geometry 1430, wherein the cell phone 1490 and thecell phone camera module 1493 is shown with an optional external lens1496. The attachment point 1415 for the reader 1410 is shown, along witha mirror 1444 and the RDT 1431 near the cell phone flash 1491.

It should be emphasized that the all five imaging/readout capabilitiesdescribed here, namely, (i) Reflection mode readout using the LEDsembedded on PCB, (ii) Reflection mode readout using cell-phone flash,(iii) Transmission mode readout using the LEDs embedded on the RDT tray(door), (iv) Transmission mode readout using the cell-phone flash withthe use of a mirror; and (v) Fluorescent mode readout using one or moreLEDs on the PCB can be implemented on the same platform with minor or nomechanical adaptation or changes. The PCB has been designed to operateat any of the readout modes. Wirelessly controlling the PCB by sendingflash pulses, the smartphone application allows the user to switchbetween readout modes or automatically chooses the readout mode based onthe RDT type.

Smartphone and Wireless Device Options

All of the currently-used, conventional reader implementations requiremultiple mechanical attachments that are physically customized to fitonto different smartphones, which may be acceptable for higher endprofessional markets, where users want to buy a complete readerinstrument and are willing to pay for the cost of the smartphone inaddition to the cost of the attachment, but it does limit the size ofthe addressable market and it is definitely too expensive for consumermarkets. Different phones can be fitted on the same reader body byhaving adjustable rails or hooks but these tend to be clumsy, expensive,and they can be misadjusted through use.

Contemplated embodiments disclosed herein implement a low-costadaptation layer by using smartphone cases. These protective cases arevery popular and sold in large quantities. Because of the huge volume,simple design, and little material they are very inexpensive ($10 to$30) and by definition they fit the smartphone 1590 perfectly. Thearchitecture of this solution is shown FIG. 15. Case 1536 is affixed tothe reader body 1510 via the attachment layer 1537, which can be screws,glue, double-sided tape, Velcro™ tape or another thin mechanicalattachment or adaptor; each with its advantages or disadvantages but allfeasible. The attachment would be permanent or semi-permanent. It wouldinitially be done in the factory and later maybe by the user. Thecassette tray 5 1521 is also shown.

With this arrangement reader body 4 can be the same for all smartphonesand rapid tests and can be made inexpensively in large quantities withhard mold injection process but, in order to be universal, it has to besomewhat larger than any contemplated smartphone. The body 1632 mustalso have an opening on the top 1634 to accommodate differentsmartphones with different positioning of the camera 1630 relative tothe main smartphone body 1690 and the flash 1637 relative to the camera1630, which is shown in FIG. 16 in an exaggerated way to illustrate thepoint.

An alternative to the use of commercially available smartphone cases isto design a universal smartphone cradle by following the same principlesas shown in FIGS. 7B and 7C, i.e. fixing the position of the smartphonein a cradle by the use of L-corner, springs, and slanted sides.

In additional contemplated embodiments, as disclosed herein, theuniversal RDT test reader can be self-contained (with an embeddeddisplay 1804) and/or have a wireless communication component that isoperatively engaged by a standalone wireless device 1805, such as asmart phone, a tablet, or another wirelessly-enabled device. Thewirelessly-enabled device has an appropriate software system,application or “app” that is designed to operatively connect with thetest reader, get and provide information from and to the reader, andmanage the information to and from the reader. The self-contained devicewith an embedded display 1901 can be used with or without anotherwirelessly-enabled device 1805 (FIG. 18). In some embodiments, thewireless communication component comprises a wireless connection that isoperatively initiated and engaged by non-optical functions. As describedherein, contemplated non-optical methods functions comprise WIFI,BLUETOOTH or Near Field Communications.

Auto-Focus Approach

The unique optical interface of the reader attachment was designed touniformly illuminate the field-of-view of an area of larger than ˜60mm×˜90 mm. This ensures that any rapid test cassette to be analyzed bythis reader will be uniformly illuminated such that the readingvariation caused by the illumination intensity is minimized, increasingthe repeatability of measurements. On the other, the digital focusing ofcell-phone camera is challenging during the image acquisition due to theneed for most uniform illumination on the RDT plane that is located only˜20-60 mm from the cell-phone camera. Live RDT image consists of onlyspatial low-frequency components at this illumination configuration,often causing the camera's auto-focus algorithm to fail. For successfulfocusing by the camera, there should be significant amount of spatialhigh-frequency components (e.g., sharp edges and transitions or lightoscillations) on the live image.

To help the camera achieve better optical focus, we first turn on thecamera's flash in burst mode to create the amount of contrast necessaryto achieve focusing on the image. This non-uniform short point sourceillumination provided by the flash generates the amount of contrastnecessary for such focus algorithms.

Once the camera has focused on the image, we are able to preserve thatfocus distance throughout the test cycle by creating a class whichimplements the Android autofocus Callback interface and setting aBoolean flag upon successful focus. We are able to obtain the pointsource illumination from the camera's flash in order to improve ourfocus while still maintaining an even, single wavelength illuminationfor the image capture. Though our solution was designed to work with acontrast detection auto-focus system, it will improve focus distancedetection for systems which use phase detection algorithms as well.

Illumination Control

Readers require sources of illumination and associated controlelectronics and battery housed outside the smartphone. In Mudanyali'sreader [18], the control is provided by the software application in thesmartphone via a cable which plugs into the smartphone micro USB powerconnector. However, many smartphones do not have the capability foroutbound control through their power connector. On the other hand, allsmartphones have an audio jack which can be used to transmit control viaan audio signal. Both approaches work but they do require externalcabling and connectors that add to the cost and reduce reliability.

Contemplated embodiments provide a wireless control connection via RFsignals, including Bluetooth, WIFI, and Near Field Communications (NFC),or using an optical signal generated by the flash in the smartphone1790. The block diagram of this approach is shown in FIG. 17. Uponusers' command to initiate the test 1715, the software (not shown)application generates an Android command that switches the flash 1767on. The burst of the flash light (not shown) is detected by a photosensor (diode or transistor) 1744 that is located on the PCB 1735,wherein the photo sensor 1744 triggers the illumination controlelectronics 1742 to activate the LEDs 1740 for RDT 1730 illumination. Asingle burst is sufficient for the usual operation where theillumination parameters are fixed and only the timing needs to becontrolled; if more complex control is desired (i.e. choice of LEDs,light level, or illumination duration), the flash 1767 can be commandedto generate a sequence of bursts that can be appropriately decoded bythe illumination control 1742. Note that this optical control method notonly avoids any external cables but it is also considerably lessexpensive than other wireless methods that might be considered. Insummary, in contemplated platforms, the flash can be used for reflectionand transmission mode illumination of RDTs as well as the wirelesscontrol of the reader attachment by the cell-phone application.

REFERENCES

The following references are referred to herein by their referencenumber. These references are incorporated herein in their entirety byreference.

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Thus, specific embodiments and methods of a universal standalone rapiddiagnostic test reader with trans-visual sensitivity have beendisclosed. It should be apparent, however, to those skilled in the artthat many more modifications besides those already described arepossible without departing from the inventive concepts herein. Theinventive subject matter, therefore, is not to be restricted except inthe spirit of the disclosure herein. Moreover, in interpreting thespecification and claims, all terms should be interpreted in thebroadest possible manner consistent with the context. In particular, theterms “comprises” and “comprising” should be interpreted as referring toelements, components, or steps in a non-exclusive manner, indicatingthat the referenced elements, components, or steps may be present, orutilized, or combined with other elements, components, or steps that arenot expressly referenced.

We claim:
 1. A universal and standalone rapid diagnostics test reader,comprising: a set of control electronics, an illumination component, animaging component, a housing component, a wireless communicationcomponent, a rapid diagnostics test component, a single universal rapiddiagnostics test tray comprising a series of cradles formed by ridges atdifferent heights, orientations, and lateral extensions, wherein thesingle tray can hold at least two different types or kinds of rapiddiagnostics test component having a shape and a size in a fixed positionrelative to the imaging component and the illumination component,wherein the single tray can only hold one test at one time and whereinthe single rapid diagnostics universal test tray can accommodatedifferent types or kinds of rapid diagnostics tests in the sameuniversal test tray without additional mechanical adaptation to eachtest before it is placed in the tray, without a mechanical adaptersurrounding each test before it is placed in the rapid diagnosticsuniversal test tray, or without additional mechanical components appliedto each test before it is placed in the rapid diagnostics universal testtray.
 2. The universal rapid diagnostics test reader of claim 1, whereinthe set of control electronics comprise processor electronics thatcontrol the illumination component, the imaging component, the wirelesscommunication component, or a combination thereof.
 3. The universalrapid diagnostics test reader of claim 1, wherein the imaging componentcomprises at least one lens, at least one image sensor, at least oneanalog to digital converter, at least one digital image processor, atleast one microprocessor, or a combination thereof.
 4. The universalrapid diagnostics test reader of claim 1, wherein the wirelesscommunication component comprises a wireless connection that isoperatively initiated and engaged by non-optical functions.
 5. Theuniversal rapid diagnostics test reader of claim 4, wherein non-opticalmethods functions comprise WIFI, BLUETOOTH or Near Field Communications.6. The universal rapid diagnostics test reader of claim 1, wherein asmartphone, tablet, laptop, computer, or another wirelessly-enableddevice operatively communicates with the universal rapid diagnosticstest reader by communicating with or engaging with the wirelesscommunication component.
 7. The universal rapid diagnostics test readerof claim 1, wherein the illumination component comprises at least onelight emitting diode.
 8. The universal rapid diagnostics test reader ofclaim 7, wherein the illumination component includes illumination of therapid diagnostics test component by the at least one light emittingdiode at the wavelength of imaging for chromatographic and colorimetricrapid diagnostic test components or at the excitation wavelength for thefluorescent rapid diagnostic test components.
 9. The universal rapiddiagnostics test reader of claim 8, wherein the illumination componentcomprises a reflection mode of operation, wherein at least onelight-emitting diode and the imaging component are on a front side ofthe rapid diagnostic test, and wherein the light-emitting diode axis isroughly perpendicular to the rapid diagnostic test component plane. 10.The universal rapid diagnostics test reader of claim 7, wherein theillumination component comprises a transmission mode of operation, andwherein the rapid diagnostic test component is between the at least onelight-emitting diode and the imaging component.
 11. The universal rapiddiagnostics test reader of claim 1, wherein the rapid diagnostic testcomponent comprises a bare strip, a strip on a translucent plasticbacking, a conventional strip packaged in a translucent plasticcassette, or a combination thereof.
 12. The universal rapid diagnosticstest reader of claim 1, wherein the rapid diagnostic test componentcomprises at least one microtiter, at least one microplate, at least oneELISA plate, at least one micro-array assay, at least one biologicaland/or chemical test, at least one other assay, or a combinationthereof.
 13. The universal rapid diagnostics test reader of claim 1,wherein the housing component is designed to enclose all components ofthe reader into a light tight enclosure and wherein the rapid diagnostictest component is illuminated only by the illumination from the readerand not illuminated by ambient light.
 14. The universal rapiddiagnostics test reader of claim 1, wherein the rapid diagnostics traysecures the rapid diagnostics test component in the tray with anL-shaped corner component.
 15. The universal rapid diagnostics testreader of claim 1, wherein the rapid diagnostic test component is heldin a fixed position in a planar direction with at least one spring. 16.The universal rapid diagnostics test reader of claim 1, wherein therapid diagnostic test component is held in a fixed position in a planardirection with at least one rack and pinion system.
 17. A universalrapid diagnostics test tray for a reader, comprising: a rapiddiagnostics test component; a tray component that is designed tooperatively couple with the reader, wherein the reader can analyze morethan one different rapid diagnostics test components, while using thesame universal rapid diagnostics test tray; and a security component,wherein the security component operatively secures the rapid diagnosticstest component in place on the tray component, wherein the traycomponent can hold at least two different types or kinds of rapiddiagnostics test component having a shape and a size in a fixed positionrelative to the imaging component and the illumination component,wherein the single tray can only hold one test at one time, and whereinthe single rapid diagnostics universal test tray comprises a series ofcradles formed by ridges at different heights, orientations, and lateralextensions and can accommodate different types or kinds of rapiddiagnostics tests in the same universal test tray without additionalmechanical adaptation to each test before it is placed in the tray,without a mechanical adapter surrounding each test before it is placedin the rapid diagnostics universal test tray, or without additionalmechanical components applied to each test before it is placed in therapid diagnostics universal test tray.
 18. The universal rapiddiagnostics test tray for a reader of claim 17, wherein the securitycomponent comprises a spring system, a rack and pinion system, or acombination thereof.